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2010 Fiscal Year Final Research Report

Improvement of protein stability for production of biofuels

Research Project

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Project/Area Number 21760638
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Biofunction/Bioprocess
Research InstitutionKobe University

Principal Investigator

TAMAKA Tsutomu  Kobe University, 自然科学系・先端融合研究環重点研究部, 助教 (90436551)

Project Period (FY) 2009 – 2010
Keywordsタンパク質 / 酵素 / 熱安定性
Research Abstract

The aim of this study is to establish enzyme-mediated protein stabilization and its application inside living cells. We have focused on Sortase, a transpeptidase. Sortase recognizes LPXTG amino acid sequences and cleaves between T and G, and then ligated with N-terminal polyglycine sequences. Our strategy is that the substrate sequences, i.e., LPXTG and GGG sequences, are introduced both N- and C-terminus of the target protein. Then Sortase allows to conjugate its N- and C-terminus and produces the circular protein. EGFP were chosen as model proteins and these substrate tags appended EGFP was successfully conjugated by Soprtase and circular EGFP was obtained. Then the thermal stability of these circular proteins was significantly improved, suggesting our strategy is very useful for protein stabilization. The optimal linker design between N-terminal and C-terminal is one of the issue for expanding versatility of this method.

  • Research Products

    (7 results)

All 2011 2010 2009

All Journal Article (2 results) Presentation (5 results)

  • [Journal Article] Site-specific tetrameric streptavidin-protein conjugation using sortase A.2011

    • Author(s)
      Matsumoto, T., Sawamoto, S., Sakamoto, T., Tanaka T., Fukuda, H., Kondo, A.
    • Journal Title

      Journal of Biotechnology 151(1-2)

      Pages: 37-42

  • [Journal Article] Enzyme-mediated site-specific antibody-protein modification using a ZZ domain as a linker.2010

    • Author(s)
      Sakamoto, T., Sawamoto, S., Tanaka T., Fukuda, H., Kondo, A.
    • Journal Title

      Bioconjugate Chemistry

      Pages: 2227-2233

  • [Presentation] Sortaseを用いたstreptavidinの新規修飾技術の開発2010

    • Author(s)
      松本拓也・田中勉・近藤昭彦
    • Organizer
      生物工学会62回大会
    • Place of Presentation
      宮崎シーガイアワールドコンベンションセンターサミット
    • Year and Date
      20101027-20101029
  • [Presentation] 酵素を用いた抗体結合ドメインー機能性タンパク質調製法の最適化2010

    • Author(s)
      坂本崇幸、田中勉、近藤昭彦
    • Organizer
      化学工学会第75年会
    • Place of Presentation
      鹿児島大学
    • Year and Date
      2010-03-20
  • [Presentation] Enzyme-mediated Antibody-Protein conjugation2009

    • Author(s)
      Takayuki Sakamoto, Tsutomu Tanaka, Akihiko Kondo.
    • Organizer
      APBioChEC 2009
    • Place of Presentation
      神戸国際会議場
    • Year and Date
      20091124-20091128
  • [Presentation] Enzyme-mediated Site-specific protein modification2009

    • Author(s)
      Tsutomu Tanaka, Takayuki Sakamoto, Kanako Wakamura, Ryosuke Takase, Takuya Matsumoto, Shiori Sawamoto, Akihiko Kondo.
    • Organizer
      APBioChEC 2009
    • Place of Presentation
      神戸国際会議場
    • Year and Date
      20091124-20091128
  • [Presentation] Sortase-mediated Site-specific protein conjugation2009

    • Author(s)
      Tsutomu Tanaka, Takayuki Sakamoto, Kanako Wakamura, Ryosuke Takase, Takuya Matsumoto, Shiori Sawamoto, Akihiko Kondo.
    • Organizer
      Enzyme Engineering XX
    • Place of Presentation
      Groningen, the Netherlands
    • Year and Date
      20090924-20090928

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Published: 2012-02-13   Modified: 2016-04-21  

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